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Molecular characterization of major and minor rDNA repeats and genetic variability assessment in different species of mahseer found in North India
Authors:Ravindra Kumar  Mamta Singh  Basdeo Kushwaha  N.S. Nagpure  Indra Mani  W.S. Lakra
Affiliation:National Bureau of Fish Genetic Resources, Canal Ring Road, P.O. Dilkusha, Lucknow 226 002, U.P., India
Abstract:Relationship among the mahseer species (Family: Cyprinidae) has long been debated in fish systematics. Present study concentrates on the nature of the phylogenetic relationship among the five mahseer species using the sequence of major ribosomal DNA (45S rDNA). We have covered rDNA sequence of approximately 5.2 kb per individual, 26.0 kb per species and 130.0 kb as a whole. We also characterized the 45S and 5S rDNA regions with respect to their nucleotide composition. For phylogenetic analyses, nucleotide sequences were divided into four datasets. First and second datasets contained 18S rDNA and ITS1 sequence, whereas third and fourth datasets consisted of ITS2 and complete 18S-ITS1-5.8S-ITS2-28S, respectively. The NJ tree was constructed for all the datasets. The mahseer species under study formed a monophyletic group well separated from the outgroup species. Similarly, the individuals of Neolissochilus hexagonolepis form monophyletic group with Tor species, indicating Neolissochilus as a sister genus of Tor. The findings from the present study provide greater insights into taxonomic status of mahseer, and set the stage for future investigations dealing with phylo-geography, taxonomy, conservation and co-evolution within this interesting and important group of fish.
Keywords:AgNO3, silver nitrate   AIC, Akaike Information Criterion   BCIP, 5-bromo-4-chloro-3&prime  -indolyphosphate p-toluidine salt   CMA3, Chromomycin A3   DNA, deoxyribonucleic acid   dNTPs, deoxynucleoside triphosphates   EMBL, European Molecular Biology Laboratory   FISH, fluorescence in situ hybridization   IE, intermediate element   ITS1, internal transcribed spacer 1   ITS2, internal transcribed spacer 2   KCl, Potassium chloride   MgCl2, Magnesium chloride   NBT, nitro-blue tetrazolium chloride   NCBI, National Center for Biotechnology Information   NJ, neighbor-joining   NH, Neolissochilus hexagonolepis   NTS, Non-transcribed spacer   PCR, polymerase chain reaction   rDNA, ribosomal deoxyribonucleic acid   SM, supplementary materials   TC, Tor chelynoides   TP, Tor putitora   TPr, Tor progeneius   TT, Tor tor   Tris&ndash  HCl, Tris hydrochloride
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